Identification of ZF-HD Gene Family in Arachis hypogaea and Analysis in Response to Abiotic Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-0806

Abstract

Abstract:

Objective To analyze the basic characteristics of the ZF-HD gene family in peanut (Arachishypogaea) and its expression patterns under various abiotic stresses, laying a foundation for revealing the role of ZF-HD in regulating abiotic stress in peanut. Method Bioinformatics methods were used to screen the peanut ZF-HD gene family members and analyze 1) their physicochemical properties, phylogenetic relationships, conserved motifs, conserved structural domains, gene structures, promoter cis-acting elements, chromosomal localization and gene covariance; 2) the expression patterns of the AhZHDs genes in different tissues and organs of peanut and in different abiotic stresses based on the transcriptome data; 3) the expressions of some members of this family in peanut following different abiotic stress treatments using RT-qPCR. The expression patterns of AhZHDs genes in different tissues and organs of peanut under different abiotic stresses were analyzed based on transcriptome data, and the expression of some members of this family in different abiotic stresses was analyzed by real-time fluorescence PCR. Result A total of 40 peanut ZF-HD genes were identified in the peanut Tifrunner genome, which were classified into six subfamilies, ZHD Ⅰ, ZHD Ⅱ, ZHD Ⅲ, ZHD Ⅳ, ZHD Ⅴ and MIF, according to their phylogenetic relationships. AhZHDs genes in the same subfamily had similar structural features, and most of them had no introns. Cis-acting element analysis indicated that the members of the peanut ZF-HD gene family were widely involved in hormone response, growth and development response, abiotic stress and light response. Chromosomal localization and covariance analysis showed that the 40 AhZHDs genes were unevenly distributed on 16 chromosomes, and that the whole genome duplication (WGD) event or fragment duplication might be the main driving force for the evolution of the AhZHDs genes. Protein interactions analysis showed that most AhZHD proteins had complex interactions with each other and might be involved in peanut development and stress response through co-regulation. Transcriptome analysis and qRT-PCR validation indicated that nine AhZHD genes (AhZHD5/9/10/17/23/29/30/32/40) showed diverse expression patterns across various peanut tissues and under stress conditions, with the majority showing significantly altered expression levels in response to abiotic stress, subcellular localization experiments showed that AhZHD5/17/29/32 function in the nucleus. Conclusion The 40 AhZHD genes differ in structure and characterization, and AhZHDs are widely involved not only in peanut growth and development and hormone signaling, but also play important regulatory roles in abiotic stresses.

Key words: peanut (A. hypogaea), ZF-HD gene, gene family, bioinformatics, tissue-specific expression, abiotic stress, expression analysis, subcellular localization

CHEN Deng-ke, LAN Gang, XIA Zhi, HOU Bao-guo, YANG Liu-liu, CAO Cai-rong, LI Peng-bo, WU Cui-cui. Identification of ZF-HD Gene Family in Arachis hypogaea and Analysis in Response to Abiotic Stress[J]. Biotechnology Bulletin, 2026, 42(4): 114-128.

Figures/Tables 12

Fig. 1 Phylogenetic tree of ZF-HD gene family in A. hypogaea, A. thaliana and O. sativa

Fig. 2 Phylogenetic relationship (A), conserved motif (B), conserved domain (C), and gene structure analysis (D) of AhZF-HD family members

Fig. 3 Analysis of cis-acting elements in the promoter regions of ZF-HD genes in A. hypogaeaA: Cis-acting elements in AhZHD family members (a: Phylogenetic relationship of AhZHD family members. b: Cis-acting elements in the AhZHD family. c: Heatmap of cis-acting elements in the AhZHD family). B: Functional categorization of cis-regulatory elements in the AhZHD family (a: Phytohormone-responsive elements. b: Elements involved in growth and development. c: Abiotic stress-responsive elements. d: Light-responsive elements). The proportion of each specific element type within the respective functional category is indicated as a percentage

Fig. 4 Distribution of AhZHDs genes on chromosomes

Fig. 5 Collinearity analysis of the ZF-HD gene family in A. hypogaea

Fig. 6 Collinearity analysis of ZF-HD genes between cultivated A. hypogaea and six other representative species

Fig. 7 Interaction network analysis of A. hypogaea ZF-HD proteins

Fig. 8 Expression analysis of A. hypogaea ZF-HD genes in different tissues

Fig. 9 Expression analysis of ZF-HD genes in A. hypogaea under high temperature, low temperature, drought, and salt stress treatments

Fig. 10 Expression differences of 9 AhZHDs genes under different tissues and organs based on RT-qPCR* and ** indicate significant differences at the level of 0.05, 0.01 respectively. The same below

Fig. 11 Expression differences of 9 AhZHDs genes under different stress treatments based on RT-qPCR

Fig. 12 Subcellular localization of AhZHD5, AhZHD17, AhZHD29 and AhZHD32 (Bar=50 μm)

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